Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1994 Nov;176(22):6992–6998. doi: 10.1128/jb.176.22.6992-6998.1994

23S rRNA domain V, a fragment that can be specifically methylated in vitro by the ErmSF (TlrA) methyltransferase.

D Kovalic 1, R B Giannattasio 1, H J Jin 1, B Weisblum 1
PMCID: PMC197072  PMID: 7961463

Abstract

The DNA sequence that encodes 23S rRNA domain V of Bacillus subtilis, nucleotides 2036 to 2672 (C. J. Green, G. C. Stewart, M. A. Hollis, B. S. Vold, and K. F. Bott, Gene 37:261-266, 1985), was cloned and used as a template from which to transcribe defined domain V RNA in vitro. The RNA transcripts served as a substrate in vitro for specific methylation of B. subtilis adenine 2085 (adenine 2058 in Escherichia coli 23S rRNA) by the ErmSF methyltransferase, an enzyme that confers resistance to the macrolide-lincosamide-streptogramin B group of antibiotics on Streptomyces fradiae NRRL 2702, the host from which it was cloned. Thus, neither RNA sequences belonging to domains other than V nor the association of 23S rRNA with ribosomal proteins is needed for the specific methylation of adenine that confers resistance to the macrolide-lincosamide-streptogramin B group of antibiotics.

Full text

PDF
6998

Images in this article

6994Image
on p.6994
6996Image
on p.6996

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Birmingham V. A., Cox K. L., Larson J. L., Fishman S. E., Hershberger C. L., Seno E. T. Cloning and expression of a tylosin resistance gene from a tylosin-producing strain of Streptomyces fradiae. Mol Gen Genet. 1986 Sep;204(3):532–539. doi: 10.1007/BF00331036. [DOI] [PubMed] [Google Scholar]
  2. Cundliffe E. How antibiotic-producing organisms avoid suicide. Annu Rev Microbiol. 1989;43:207–233. doi: 10.1146/annurev.mi.43.100189.001231. [DOI] [PubMed] [Google Scholar]
  3. Denoya C., Dubnau D. Mono- and dimethylating activities and kinetic studies of the ermC 23 S rRNA methyltransferase. J Biol Chem. 1989 Feb 15;264(5):2615–2624. [PubMed] [Google Scholar]
  4. Douthwaite S., Powers T., Lee J. Y., Noller H. F. Defining the structural requirements for a helix in 23 S ribosomal RNA that confers erythromycin resistance. J Mol Biol. 1989 Oct 20;209(4):655–665. doi: 10.1016/0022-2836(89)93000-3. [DOI] [PubMed] [Google Scholar]
  5. Douthwaite S., Prince J. B., Noller H. F. Evidence for functional interaction between domains II and V of 23S ribosomal RNA from an erythromycin-resistant mutant. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8330–8334. doi: 10.1073/pnas.82.24.8330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Green C. J., Stewart G. C., Hollis M. A., Vold B. S., Bott K. F. Nucleotide sequence of the Bacillus subtilis ribosomal RNA operon, rrnB. Gene. 1985;37(1-3):261–266. doi: 10.1016/0378-1119(85)90281-1. [DOI] [PubMed] [Google Scholar]
  7. Gutell R. R., Fox G. E. A compilation of large subunit RNA sequences presented in a structural format. Nucleic Acids Res. 1988;16 (Suppl):r175–r269. doi: 10.1093/nar/16.suppl.r175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Iwanami Y., Brown G. M. Methylated bases of ribosomal ribonucleic acid from HeLa cells. Arch Biochem Biophys. 1968 Jul;126(1):8–15. doi: 10.1016/0003-9861(68)90553-5. [DOI] [PubMed] [Google Scholar]
  9. Kamimiya S., Weisblum B. Translational attenuation control of ermSF, an inducible resistance determinant encoding rRNA N-methyltransferase from Streptomyces fradiae. J Bacteriol. 1988 Apr;170(4):1800–1811. doi: 10.1128/jb.170.4.1800-1811.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lai C. J., Dahlberg J. E., Weisblum B. Structure of an inducibly methylatable nucleotide sequence in 23S ribosomal ribonucleic acid from erythromycin-resistant Staphylococcus aureus. Biochemistry. 1973 Jan 30;12(3):457–460. doi: 10.1021/bi00727a015. [DOI] [PubMed] [Google Scholar]
  11. Lai C. J., Weisblum B. Altered methylation of ribosomal RNA in an erythromycin-resistant strain of Staphylococcus aureus. Proc Natl Acad Sci U S A. 1971 Apr;68(4):856–860. doi: 10.1073/pnas.68.4.856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lai C. J., Weisblum B., Fahnestock S. R., Nomura M. Alteration of 23 S ribosomal RNA and erythromycin-induced resistance to lincomycin and spiramycin in Staphylococcus aureus. J Mol Biol. 1973 Feb 15;74(1):67–72. doi: 10.1016/0022-2836(73)90355-0. [DOI] [PubMed] [Google Scholar]
  13. Mullis K. B., Faloona F. A. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. doi: 10.1016/0076-6879(87)55023-6. [DOI] [PubMed] [Google Scholar]
  14. Noller H. F., Hoffarth V., Zimniak L. Unusual resistance of peptidyl transferase to protein extraction procedures. Science. 1992 Jun 5;256(5062):1416–1419. doi: 10.1126/science.1604315. [DOI] [PubMed] [Google Scholar]
  15. Noller H. F. Peptidyl transferase: protein, ribonucleoprotein, or RNA? J Bacteriol. 1993 Sep;175(17):5297–5300. doi: 10.1128/jb.175.17.5297-5300.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Noller H. F. Ribosomal RNA and translation. Annu Rev Biochem. 1991;60:191–227. doi: 10.1146/annurev.bi.60.070191.001203. [DOI] [PubMed] [Google Scholar]
  17. Porath J., Carlsson J., Olsson I., Belfrage G. Metal chelate affinity chromatography, a new approach to protein fractionation. Nature. 1975 Dec 18;258(5536):598–599. doi: 10.1038/258598a0. [DOI] [PubMed] [Google Scholar]
  18. Rosenberg A. H., Lade B. N., Chui D. S., Lin S. W., Dunn J. J., Studier F. W. Vectors for selective expression of cloned DNAs by T7 RNA polymerase. Gene. 1987;56(1):125–135. doi: 10.1016/0378-1119(87)90165-x. [DOI] [PubMed] [Google Scholar]
  19. Shivakumar A. G., Dubnau D. Characterization of a plasmid-specified ribosome methylase associated with macrolide resistance. Nucleic Acids Res. 1981 Jun 11;9(11):2549–2562. doi: 10.1093/nar/9.11.2549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sigmund C. D., Ettayebi M., Borden A., Morgan E. A. Antibiotic resistance mutations in ribosomal RNA genes of Escherichia coli. Methods Enzymol. 1988;164:673–690. doi: 10.1016/s0076-6879(88)64077-8. [DOI] [PubMed] [Google Scholar]
  21. Skinner R., Cundliffe E., Schmidt F. J. Site of action of a ribosomal RNA methylase responsible for resistance to erythromycin and other antibiotics. J Biol Chem. 1983 Oct 25;258(20):12702–12706. [PubMed] [Google Scholar]
  22. Smith J. E., Cooperman B. S., Mitchell P. Methylation sites in Escherichia coli ribosomal RNA: localization and identification of four new sites of methylation in 23S rRNA. Biochemistry. 1992 Nov 10;31(44):10825–10834. doi: 10.1021/bi00159a025. [DOI] [PubMed] [Google Scholar]
  23. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  24. Su S. L., Dubnau D. Binding of Bacillus subtilis ermC' methyltransferase to 23S rRNA. Biochemistry. 1990 Jun 26;29(25):6033–6042. doi: 10.1021/bi00477a022. [DOI] [PubMed] [Google Scholar]
  25. Weisblum B., Graham M. Y., Gryczan T., Dubnau D. Plasmid copy number control: isolation and characterization of high-copy-number mutants of plasmid pE194. J Bacteriol. 1979 Jan;137(1):635–643. doi: 10.1128/jb.137.1.635-643.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Zalacain M., Cundliffe E. Methylation of 23S rRNA caused by tlrA (ermSF), a tylosin resistance determinant from Streptomyces fradiae. J Bacteriol. 1989 Aug;171(8):4254–4260. doi: 10.1128/jb.171.8.4254-4260.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES